CN110006873A - Environmental pollutant detection method based on three-dimensional micro-nano structure enhancing Raman spectrum - Google Patents
Environmental pollutant detection method based on three-dimensional micro-nano structure enhancing Raman spectrum Download PDFInfo
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- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
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Abstract
The present invention provides a kind of environmental pollutant detection methods based on three-dimensional micro-nano structure enhancing Raman spectrum, include the following steps: that copper particle is impregnated 5-15min using dilution heat of sulfuric acid first by (1), soaked copper particle is passed through in PDMS microchannel, and silver nitrate solution is injected with the rate of 10 μ L/min, reaction time is 5-15min, copper particle is finally rinsed using ethyl alcohol, until the liquid wash on copper surface is clean, i.e. acquisition three-dimensional micro-nano structure SERS substrate；(2) sample to be tested is configured to solution, and is passed through in PDMS microchannel with the rate of 10 μ L/min；(3) micro-fluidic system is placed on the objective table of confocal detection platform, adjusts microcobjective at a distance from sample, laser spot is focused on sample, the raman spectral signal of sample to be tested is acquired.The detection method high sensitivity, it is portable miniaturized, and the SERS substrate prepared has filterability and identity to cancer cell.
Description
Technical field
The invention belongs to Surface enhanced Raman spectroscopy analysis technical fields, and in particular to one kind is increased based on three-dimensional micro-nano structure
The environmental pollutant detection method of strong Raman spectrum.
Background technique
With the continuous deepening of research, the application of Surface enhanced Raman spectroscopy (SERS) has spread over every field.?
In these applications, the SERS test in conjunction with microfluidic device has shown in traditional macro environment some apparent excellent
Gesture.Currently, being all by most of enhancing substrates that metal Nano structure forms in microfluidic surface enhancing raman study
It directly prepares in the channel.For example, Kang et al. reports a kind of photoinduction synthetic method, it is used to prepare to be embedded in and is integrated into
The silver nano-grain on the zinc oxide nanowire surface of microfluidic system；Bai et al. is prepared for a kind of wet based on femtosecond laser auxiliary
The three-dimensional microfluidic SERS system of method etching；When Leem et al. heats substrate with polyol process, the shape directly in microfluidic channel
At Ag films.In addition, other methods, such as photo-reduction and photo-thermal effect have also been used to manufacture enhancing in micro-fluidic system
Substrate.
However, most of is all directly to prepare two dimension enhancing base in microchannel in these two-dimensional micro-fluidic systems
Bottom, in the liquid of flowing, since the molecule of low concentration flows in several hundred microns of channel, it is difficult to be attached to substrate table
Face, thus testing molecule is difficult to reach metal surface enhancing hot spot (≤100nm), leads to the Raman spectrum detection effect of molecule not
It is good, certain difficulty is brought to SERS detection.There is researcher to synthesize three-dimensional S ERS substrate, mainly three-dimensional structure in microchannel
Particle have high surface area to reach the hot spot of metal structure surface, help to enhance SERS signal, and realize to be measured point
The detection of son.But these method equipment are expensive, preparation process is complicated, time-consuming, testing cost is high.
Therefore, research and develop the convenient method that a kind of SERS substrate is used to improve SERS signal, thus accurately analysis it is different to
The Raman spectrum of sample is to solve time-consuming one of the critical issue with reduction equipment cost expense of miniflow SERS test.
Summary of the invention
Aiming at the above shortcomings existing in the prior art, the purpose of the present invention is to provide one kind to be based on three-dimensional micro-nano structure
Enhance the environmental pollutant detection method of Raman spectrum, the detection method high sensitivity is portable miniaturized, and the SERS prepared
Substrate has filterability and identity to cancer cell, can satisfy the new demand of the application aspects such as food safety assessment.
To achieve the above object, the invention provides the following technical scheme: a kind of enhance Raman light based on three-dimensional micro-nano structure
The environmental pollutant detection method of spectrum, includes the following steps:
(1) copper particle is impregnated by 5-15min using dilution heat of sulfuric acid first, soaked copper particle is passed through PDMS miniflow
In pipeline, the amount for being passed through copper particle is the 1/3-1/2 of microchannel volume, and molten with the rate of 10 μ L/min injection silver nitrate
Liquid, reaction time 5-15min finally rinse copper particle using ethyl alcohol, until the liquid wash on copper surface is clean, that is, obtain
Three-dimensional micro-nano structure SERS substrate；
(2) sample to be tested is configured to solution, and is passed through in PDMS microchannel with the rate of 10 μ L/min, be passed through the time
For 10min；
(3) micro-fluidic system is placed on the objective table of confocal detection platform, adjust microcobjective and sample away from
From laser spot is focused on sample, the raman spectral signal of sample to be tested is acquired.
Further, the dilution heat of sulfuric acid that the dilute sulfuric acid used in step (1) is 1.0-3.0% for mass fraction, the nitre
The concentration of sour silver solution is 1-10mM.Use the oxidation that the purpose that dilute sulfuric acid impregnates copper particle is to remove copper particle surface
Layer.
Further, the condition of sample to be tested raman spectral signal is acquired in step (3) are as follows: optical maser wavelength 633nm, laser
Power is 10mW, acquisition time 20s.
It is compared with the prior art, the invention has the following beneficial effects:
Environmental pollutant detection method high sensitivity provided by the invention based on three-dimensional micro-nano structure enhancing Raman spectrum,
Can be achieved portable inspectiont, while prepare SRES substrate method have preparation time it is short, at low cost, easy to operate, it can be achieved that
The characteristics of SERS of different testing molecules is detected, and the SERS substrate prepared has filterability and identity to cancer cell, it can
To meet the new demand of the application aspects such as food safety assessment.
Detailed description of the invention
Fig. 1 is the SEM figure of three-dimensional micro-nano structure SERS substrate synthesis front and back；
Fig. 2 is the Raman spectrogram of methylene blue (MB)；
Fig. 3 is the Raman spectrogram of urea；
Fig. 4 is the Raman spectrogram of melamine；
Fig. 5 is the Raman spectrum of 231 breast cancer cell of H1299 lung carcinoma cell and MDA-MB；
Fig. 6 is the SERS substrate right side, a left side, bottom, characteristic peak is 1618cm at the four direction of top-1Raman spectrogram；
Fig. 7 is the simulation drawing that microchannel passes in and out mouth cells.
Specific embodiment
A kind of environmental pollutant detection method based on three-dimensional micro-nano structure enhancing Raman spectrum, includes the following steps:
(1) it uses mass fraction that copper particle is impregnated 10min for the dilution heat of sulfuric acid of 1.0-3.0% first, will impregnate
Copper particle be passed through in PDMS microchannel, the amount for being passed through copper particle is the 1/3-1/2 of microchannel volume, and with 10 μ L/min
Rate implantation concentration be 1-4mM silver nitrate solution, reaction time 5-15min, finally using ethyl alcohol rinse copper particle, until
Until the liquid wash of copper particle surface is clean, that is, obtain three-dimensional micro-nano structure SERS substrate；
(2) sample to be tested is configured to solution, and is passed through in PDMS microchannel with the rate of 10 μ L/min, be passed through the time
For 10min；
(3) micro-fluidic system is placed on the objective table of confocal detection platform, adjust microcobjective and sample away from
From, laser spot is focused on sample, acquires the raman spectral signal of sample to be tested, the condition of Raman signal acquisition are as follows: swash
Light wave a length of 633nm, laser power 10mW, acquisition time 20s.
Fig. 1 is SEM figure, wherein figure (a) is the SEM figure before the synthesis of SERS substrate, it can be seen that the copper before synthesis
Grain surface is more smooth；Scheming (b) is the SEM figure after the synthesis of SERS substrate, since the Ag that displacement reaction generates is wrapped in Cu particle table
Face, to form the copper Ag doping three-dimensional wiener structure with micron order gap.
Detection method of the invention can detect methylene blue (MB) or urea or melamine solution, also can be used
In filtering and identification to cancer cell.
Combined with specific embodiments below, the method for the present invention is described in detail.The microchannel used in the present invention
Input end dimension is 600*150*1000 μm (wide * high * long), and outlet end is 30*150*1000 μm.
Embodiment 1
The Raman spectrum of methylene blue (MB) is detected, specific to detect as follows:
(1) use mass fraction that copper particle is impregnated 10min for 2.0% dilution heat of sulfuric acid first, by soaked copper
Particle is passed through in PDMS microchannel, and the amount for being passed through copper particle is the 1/3-1/2 of microchannel volume, and with the speed of 10 μ L/min
Rate implantation concentration is 1mM silver nitrate solution, and reaction time 10min finally rinses copper particle using ethyl alcohol, until copper particle table
Until the liquid wash in face is clean, that is, obtain three-dimensional micro-nano structure SERS substrate；
(2) MB is configured to concentration is 10-4M、10-5M、10-6M、10-7M and 10-8The aqueous solution of M, and with 10 μ L/min's
Above-mentioned solution is passed through in PDMS microchannel by rate, and being passed through the time is 10min；Concrete operations are by needle tubing by above-mentioned difference
The solution of concentration is passed through in pipeline, is passed through by the sequence by low concentration to high concentration；
(3) micro-fluidic system is placed on the objective table of confocal detection platform, adjust microcobjective and sample away from
From, laser spot is focused on sample, acquires the raman spectral signal of sample to be tested, the condition of Raman signal acquisition are as follows: swash
Light wave a length of 633nm, laser power 10mW, acquisition time 20s.As a result as shown in Fig. 2, it can be seen from the figure that using
Detection method of the invention is in 1618cm-1Place can obtain the raman characteristic peak of the MB of various concentration, and detection side of the invention
It is 10 that method, which can detecte concentration,-7The raman characteristic peak of the MB of M, this illustrates detection method high sensitivity of the invention.
Embodiment 2
The Raman spectrum of urea is detected, specific to detect as follows:
(1) use mass fraction that copper particle is impregnated 10min for 1.0% dilution heat of sulfuric acid first, by soaked copper
Particle is passed through in PDMS microchannel, and the amount for being passed through copper particle is the 1/3-1/2 of microchannel volume, and with the speed of 10 μ L/min
Rate implantation concentration is 2mM silver nitrate solution, and reaction time 5min finally rinses copper particle using ethyl alcohol, until copper particle table
Until the liquid wash in face is clean, that is, obtain three-dimensional micro-nano structure SERS substrate；
(2) it takes certain deionized water to be heated to 90 DEG C, and urea is added into deionized water, being configured to concentration is 10- 4M、10-5M、10-6M、10-7M and 10-8The aqueous solution of M, and above-mentioned solution is passed through by PDMS microchannel with the rate of 10 μ L/min
In, being passed through the time is 10min；Concrete operations are that the solution of above-mentioned various concentration is passed through in pipeline by needle tubing, by by low dense
The sequence for spending high concentration is passed through；
(3) micro-fluidic system is placed on the objective table of confocal detection platform, adjust microcobjective and sample away from
From, laser spot is focused on sample, acquires the raman spectral signal of sample to be tested, the condition of Raman signal acquisition are as follows: swash
Light wave a length of 633nm, laser power 10mW, acquisition time 20s.As a result as shown in figure 3, it can be seen from the figure that using
Detection method of the invention is in 965cm-1Place can obtain the Raman signatures spectrum peak of the urea of various concentration.
Embodiment 3
The Raman spectrum of melamine is detected, specific to detect as follows:
(1) use mass fraction that copper particle is impregnated 10min for 3.0% dilution heat of sulfuric acid first, by soaked copper
Particle is passed through in PDMS microchannel, and the amount for being passed through copper particle is the 1/3-1/2 of microchannel volume, and with the speed of 10 μ L/min
Rate implantation concentration is 4mM silver nitrate solution, and reaction time 150min finally rinses copper particle using ethyl alcohol, until copper particle
Until the liquid wash on surface is clean, that is, obtain three-dimensional micro-nano structure SERS substrate；
(2) it takes certain deionized water to be heated to 90 DEG C, and melamine is added into deionized water, being configured to concentration is
10-5M、10-6M、10-7M and 10-8The aqueous solution of M, and above-mentioned solution is passed through in PDMS microchannel with the rate of 10 μ L/min,
Being passed through the time is 10min；Concrete operations are that the solution of above-mentioned various concentration is passed through in pipeline by needle tubing, by by low concentration to
The sequence of high concentration is passed through；
(3) micro-fluidic system is placed on the objective table of confocal detection platform, adjust microcobjective and sample away from
From, laser spot is focused on sample, acquires the raman spectral signal of sample to be tested, the condition of Raman signal acquisition are as follows: swash
Light wave a length of 633nm, laser power 10mW, acquisition time 20s.As a result as shown in figure 4, it can be seen from the figure that using
Detection method of the invention is in 953cm-1Place can obtain the Raman signatures spectrum peak of the melamine of various concentration.
Embodiment 4
The Raman spectrum of 231 breast cancer cell of H1299 lung carcinoma cell in culture solution and MDA-MB is detected, specifically
It detects as follows:
(1) use mass fraction that copper particle is impregnated 10min for 2.0% dilution heat of sulfuric acid first, by soaked copper
Particle is passed through in PDMS microchannel, and the amount for being passed through copper particle is the 1/3-1/2 of microchannel volume, and with the speed of 10 μ L/min
Rate implantation concentration is 10mM silver nitrate solution, and reaction time 10min finally rinses copper particle using ethyl alcohol, until copper particle
Until the liquid wash on surface is clean, that is, obtain three-dimensional micro-nano structure SERS substrate；
(2) by H1299 lung carcinoma cell and 231 breast cancer cell of MDA-MB in 37 DEG C, 5%CO2Incubator in cultivate,
The cancer cell containing culture solution is passed through in PDMS microchannel with the rate of 10 μ L/min, being passed through the time is 10min；
(3) and by PBS solution it is passed through pipe flushing cancer cell, collection device exit solution carries out raman spectral signal
Acquisition, the condition of Raman signal acquisition are as follows: optical maser wavelength 633nm, laser power 10mW, acquisition time 20s.As a result such as
Shown in Fig. 5, from a figure and b figure as can be seen that using detection method of the invention in 1386cm-1Place and 670cm-1、994cm-1
Place can obtain the Raman signatures spectrum peak of 231 breast cancer cell of H1299 lung carcinoma cell and MDA-MB respectively.
Embodiment 5
Verify the stability of SERS substrate prepared by the present invention
By taking Raman spectrum of the embodiment 1 to MB detects as an example, micro-fluidic system is placed in confocal detection platform
Objective table on, respectively from microchannel right side, left side, bottom and top four direction acquisition MB raman spectral signal.Its
It is middle to choose 6 peak intensity positions as 1618cm-1The characteristic peak measured value at place, for analyzing the stability of SERS substrate.As a result such as Fig. 5
It is shown, the raman spectral signal of MB as can be seen from the figure can be collected in the right side of pipeline, left side, bottom and top,
In, the signal acquired from duct bottom is most strong, followed by top, left side, right side；This illustrates three-D micro-nano knot prepared by the present invention
The three-dimensional stability of structure SERS substrate is good, can be from either SERS substrate to acquisition signal.
Embodiment 6
Three-dimensional micro-nano structure SERS substrate prepared by the present invention is verified to the filterability and recognition performance of cell
Cell is introduced by import, and is contacted with three-dimensional micro-nano structure SERS substrate, it is more in the case where directly filtering
Remaining liquid will be flowed out by outlet, and cell accumulation is in pipeline, in the cell-free outflow in exit.With circulating repetition different in flow rate
Several times, outlet port does not occur cell, it was demonstrated that this three-dimensional microfluidic systems has outstanding strainability.As a result such as table 1
It is shown:
The lower inlet and outlet cell number comparison different in flow rate of table 1
Wherein, the input end dimension of used microchannel is 600*150*1000 μm (wide * high * long), and outlet end is
30*150*1000μm。
As can be seen from the table, under different in flow rate, when impurity and cell simultaneously from the outlet of three-dimensional microfluidic systems into
Enter, by three-dimensional S ERS substrate, cell finally is not detected in exit, this explanation uses three-dimensional S ERS substrate system of the invention
Standby microfluidic devices have filterability and recognition performance to cell.
Finally, it is stated that the above examples are only used to illustrate the technical scheme of the present invention and are not limiting, although referring to compared with
Good embodiment describes the invention in detail, those skilled in the art should understand that, it can be to skill of the invention
Art scheme is modified or replaced equivalently, and without departing from the objective and range of technical solution of the present invention, should all be covered at this
In the scope of the claims of invention.
Claims (3)
1. a kind of environmental pollutant detection method based on three-dimensional micro-nano structure enhancing Raman spectrum, which is characterized in that including such as
Lower step:
(1) copper particle is impregnated by 5-15min using dilution heat of sulfuric acid first, soaked copper particle is passed through PDMS microchannel
In, the amount for being passed through copper particle is the 1/3-1/2 of microchannel volume, and injects silver nitrate solution with the rate of 10 μ L/min, instead
It is 5-15min between seasonable, copper particle is finally rinsed using ethyl alcohol, until the liquid wash on copper surface is clean, i.e. acquisition is three-dimensional
Micro-nano structure SERS substrate；
(2) sample to be tested is configured to solution, and is passed through in PDMS microchannel with the rate of 10 μ L/min, being passed through the time is
10min；
(3) micro-fluidic system is placed on the objective table of confocal detection platform, adjusts microcobjective at a distance from sample, it will
Laser spot focuses on sample, acquires the raman spectral signal of sample to be tested.
2. the environmental pollutant detection method according to claim 1 based on three-dimensional micro-nano structure enhancing Raman spectrum,
It is characterized in that, the dilution heat of sulfuric acid that the dilute sulfuric acid used in step (1) is 1.0-3.0% for mass fraction, the silver nitrate is molten
The concentration of liquid is 1-10mM.
3. the environmental pollutant detection method according to claim 1 based on three-dimensional micro-nano structure enhancing Raman spectrum,
It is characterized in that, the condition of acquisition sample to be tested raman spectral signal in step (3) are as follows: optical maser wavelength 633nm, laser power are
10mW, acquisition time 20s.
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| CN113624742A (en) * | 2021-08-31 | 2021-11-09 | 重庆大学 | Hydrogen enhanced Raman spectroscopy detection method for sulfhydryl gas in plasmon MOF |
| CN113877643A (en) * | 2021-09-18 | 2022-01-04 | 重庆市益康环保工程有限公司 | Cellulose 3D enhanced Raman spectrum microfluidic chip for detecting water pollution and application thereof |
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